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in vitro neuraminidase inhibitory activity of four neuraminidase inhibitors against influenza virus isolates in the 2011-2012 season in japan.the neuraminidase inhibitors oseltamivir phosphate (tamiflu®), zanamivir (relenza®), laninamivir octanoate (inavir®), and peramivir (rapiacta®) have been available for the treatment of influenza in japan since 2010. to assess the extent of viral resistance, we measured the 50% inhibitory concentration (ic₅₀) of each drug for influenza virus isolates from the 2011-2012 influenza season. specimens were obtained from patients prior to treatment. viral isolation was done using madine-darby canine ki ...201424560563
h3n2 virus as causative agent of ards requiring extracorporeal membrane oxygenation support.pandemic influenza virus a(h1n1) 2009 was associated with a higher risk of viral pneumonia in comparison with seasonal influenza viruses. the influenza season 2011-2012 was characterized by the prevalent circulation of influenza a(h3n2) viruses. whereas most h3n2 patients experienced mild, self-limited influenza-like illness, some patients were at increased risk for influenza complications because of age or underlying medical conditions. cases presented were patients admitted to the intensive ca ...201424527039
phylogenetic analysis of a swine influenza a(h3n2) virus isolated in korea in 2012.influenza a virus (iav) can infect avian and mammalian species, including humans. the genome nature of iavs may contribute to viral adaptation in different animal hosts, resulting in gene reassortment and the reproduction of variants with optimal fitness. as seen again in the 2009 swine-origin influenza a h1n1 pandemic, pigs are known to be susceptible to swine, avian, and human iavs and can serve as a 'mixing vessel' for the generation of novel iav variants. to this end, the emergence of swine ...201424523938
tmprss2 is a host factor that is essential for pneumotropism and pathogenicity of h7n9 influenza a virus in mice.cleavage of the hemagglutinin (ha) by host proteases is essential for the infectivity of influenza viruses. here, we analyzed the role of the serine protease tmprss2, which activates ha in the human respiratory tract, in pathogenesis in a mouse model. replication of the human h7n9 isolate a/anhui/1/13 and of human h1n1 and h3n2 viruses was compared in tmprss2 knockout (tmprss2(-/-)) and wild-type (wt) mice. knockout of tmprss2 expression inhibited h7n9 influenza virus replication in explants of ...201424522916
prospective surveillance and molecular characterization of seasonal influenza in a university cohort in singapore.southeast asia is believed to be a potential locus for the emergence of novel influenza strains, and therefore accurate sentinel surveillance in the region is critical. limited information exists on sentinel surveillance of influenza-like illness (ili) in young adults in singapore in a university campus setting. the objective of the present study was to determine the proportion of ili caused by influenza a and b viruses in a university cohort in singapore.201424520371
influenza a virus infections in marine mammals and terrestrial carnivores.influenza a viruses (iav), members of the orthomyxoviridae, cover a wide host spectrum comprising a plethora of avian and, in comparison, a few mammalian species. the viral reservoir and gene pool are kept in metapopulations of aquatic wild birds. the mammalian-adapted iavs originally arose by transspecies transmission from avian sources. in swine, horse and man, species-adapted iav lineages circulate independently of the avian reservoir and cause predominantly respiratory disease of highly vari ...201424511825
integrating influenza antigenic dynamics with molecular evolution.influenza viruses undergo continual antigenic evolution allowing mutant viruses to evade host immunity acquired to previous virus strains. antigenic phenotype is often assessed through pairwise measurement of cross-reactivity between influenza strains using the hemagglutination inhibition (hi) assay. here, we extend previous approaches to antigenic cartography, and simultaneously characterize antigenic and genetic evolution by modeling the diffusion of antigenic phenotype over a shared virus phy ...201424497547
differences in antibody responses between trivalent inactivated influenza vaccine and live attenuated influenza vaccine correlate with the kinetics and magnitude of interferon signaling in children.live attenuated influenza vaccine (laiv) and trivalent inactivated influenza vaccine (tiv) are effective for prevention of influenza virus infection in children, but the mechanisms associated with protection are not well defined.201424495909
a new class of synthetic anti-lipopolysaccharide peptides inhibits influenza a virus replication by blocking cellular attachment.influenza a viruses are a continuous threat to human health as illustrated by the 2009 h1n1 pandemic. since circulating influenza virus strains become increasingly resistant against currently available drugs, the development of novel antivirals is urgently needed. here, we have evaluated a recently described new class of broad-spectrum antiviral peptides (synthetic anti-lipopolysaccharide peptides; salps) for their potential to inhibit influenza virus replication in vitro and in vivo. we found t ...201424486207
genetic characterization of influenza a viruses circulating in pigs and isolated in north-east spain during the period 2006-2007.swine influenza virus is one of the most important pathogens involved in the swine respiratory disease complex. recent serological surveys showed a high prevalence of swine influenza strains belonging to the h1n1, h1n2 and h3n2 subtypes circulating in pigs in spain. however, little is known about their genome sequence. five swine influenza strains were isolated from some unrelated outbreaks occurred during 2006-2007, and their complete genome sequences were determined. phylogenetic analysis reve ...201424461956
recombinant influenza h7 hemagglutinins induce lower neutralizing antibody titers in mice than do seasonal hemagglutinins.vaccines against avian influenza viruses often require high hemagglutinin (ha) doses or adjuvants to achieve serological titers associated with protection against disease. in particular, viruses of the h7 subtype frequently do not induce strong antibody responses following immunization.201425213778
effectiveness of post-exposition prophylaxis with oseltamivir in nursing homes: a randomised controlled trial over four seasons.oseltamivir has been registered for use as post-exposition prophylaxis (pep) following exposure to influenza, based on studies among healthy adults. effectiveness among frail elderly nursing home populations still needs to be properly assessed.201425210532
characterization of a broadly neutralizing monoclonal antibody that targets the fusion domain of group 2 influenza a virus hemagglutinin.due to continuous changes to its antigenic regions, influenza viruses can evade immune detection and cause a significant amount of morbidity and mortality around the world. influenza vaccinations can protect against disease but must be annually reformulated to match the current circulating strains. in the development of a broad-spectrum influenza vaccine, the elucidation of conserved epitopes is paramount. to this end, we designed an immunization strategy in mice to boost the humoral response ag ...201425210195
emerging viral respiratory tract infections--environmental risk factors and transmission.the past decade has seen the emergence of several novel viruses that cause respiratory tract infections in human beings, including middle east respiratory syndrome coronavirus (mers-cov) in saudi arabia, an h7n9 influenza a virus in eastern china, a swine-like influenza h3n2 variant virus in the usa, and a human adenovirus 14p1 also in the usa. mers-cov and h7n9 viruses are still a major worldwide public health concern. the pathogenesis and mode of transmission of mers-cov and h7n9 influenza a v ...201425189350
generation and characterization of influenza a viruses with altered polymerase fidelity.genetic diversity of influenza a viruses (iav) acquired through the error-prone rna-dependent rna polymerase (rdrp) or through genetic reassortment enables perpetuation of iav in humans through epidemics or pandemics. here, to assess the biological significance of genetic diversity acquired through rdrp, we characterize an iav fidelity variant derived from passaging a seasonal h3n2 virus in the presence of ribavirin, a purine analogue that increases guanosine-to-adenosine mutations. we demonstra ...201425183443
a population prevalence study on influenza infection in dogs in southern italy.despite several data on influenza infection in dogs, the first natural outbreak of canine influenza virus, closely related to h3n8 equine subtype, dates back to 2004 in florida. subsequent studies highlighted the role of dogs in adaptation of h5n1 to mammals and the susceptibility of dogs to different subtypes of influenza. a prevalence study was carried out on 562 sera collected from pet and kennel dogs in the south of italy. a c-elisa test was employed and c-elisa-positive, c-elisa-doubtful an ...201425180843
infection of influenza virus neuraminidase-vaccinated mice with homologous influenza virus leads to strong protection against heterologous influenza viruses.vaccination is the best measure to prevent influenza pandemics. here, we studied the protective effect against heterologous influenza viruses, including a/reassortant/nymc x-179a (ph1n1), a/chicken/henan/12/2004 (h5n1), a/chicken/jiangsu/7/2002 (h9n2) and a/guizhou/54/89×a/pr/8/34 (a/guizhou-x) (h3n2), in mice first vaccinated with a dna vaccine of haemagglutinin (ha) or neuraminidase (na) of a/pr/8/34 (pr8) and then infected with the homologous virus. we showed that pr8 ha or na vaccination bot ...201425170051
influenza polymerase encoding mrnas utilize atypical mrna nuclear export.influenza is a segmented negative strand rna virus. each rna segment is encapsulated by influenza nucleoprotein and bound by the viral rna dependent rna polymerase (rdrp) to form viral ribonucleoproteins responsible for rna synthesis in the nucleus of the host cell. influenza transcription results in spliced mrnas (m2 and ns2), intron-containing mrnas (m1 and ns1), and intron-less mrnas (ha, na, np, pb1, pb2, and pa), all of which undergo nuclear export into the cytoplasm for translation. most c ...201425168591
early activation of map kinases by influenza a virus x-31 in murine macrophage cell lines.early molecular responses to influenza a (flua) virus strain a/x-31 h3n2 in macrophages were explored using j774.a1 and raw 264.7 murine cell lines. nf-kappa b (nfκb) was reported to be central to flua host-response in other cell types. our data showed that flua activation of the classical nfκb dependent pathway in these macrophages was minimal. regulator proteins, ikappab-alpha and -beta (iκbα, iκbβ), showed limited degradation peaking at 2 h post flua exposure and p65 was not observed to trans ...201425166426
deep sequencing reveals the eight facets of the influenza a/hongkong/1/1968 (h3n2) virus cap-snatching process.the influenza a virus rna polymerase cleaves the 5' end of host pre-mrnas and uses the capped rna fragments as primers for viral mrna synthesis. we performed deep sequencing of the 5' ends of viral mrnas from all genome segments transcribed in both human (a549) and mouse (m-1) cells infected with the influenza a/hongkong/1/1968 (h3n2) virus. in addition to information on rna motifs present, our results indicate that the host primers are divergent between the viral transcripts. we observed differ ...201425154590
swine-to-human transmission of influenza a(h3n2) virus at agricultural fairs, ohio, usa, 2012.agricultural fairs provide an opportunity for bidirectional transmission of influenza a viruses. we sought to determine influenza a virus activity among swine at fairs in the united states. as part of an ongoing active influenza a virus surveillance project, nasal swab samples were collected from exhibition swine at 40 selected ohio agricultural fairs during 2012. influenza a(h3n2) virus was isolated from swine at 10 of the fairs. according to a concurrent public health investigation, 7 of the 1 ...201425148572
characterization of variations in pb2, ns1, m, neuraminidase and hemagglutinin of influenza a (h3n2) viruses in iran.in the influenza a viruses, neuraminidase (na), hemagglutinin (ha), pb2, ns1 and m are responsible for the disease pathogenicity. the mechanism of pathogenicity differs among these viruses. binding of host proteases by the viral na, sequence of ha in the cleavage and receptor-binding sites, number of oligosaccharide side chains of ha, shortening of na, and substitutions in pb2, ns1 and m genes, all have been suggested as molecular correlates of pathogenicity of influenza viruses.201425147684
computational prediction of vaccine strains for human influenza a (h3n2) viruses.human influenza a viruses are rapidly evolving pathogens that cause substantial morbidity and mortality in seasonal epidemics around the globe. to ensure continued protection, the strains used for the production of the seasonal influenza vaccine have to be regularly updated, which involves data collection and analysis by numerous experts worldwide. computer-guided analysis is becoming increasingly important in this problem due to the vast amounts of generated data. we here describe a computation ...201425122778
[genetic characteristics of influenza a/h3n2 virus neuraminidase gene: a survey from 2010 to 2012 in qinghai province, china].this study aims to perform a survey of genetic variation in neuraminidase (na) gene of influenza a/h3n2 virus, as well as related resistance to na inhibitors, in qinghai province of china, 2010 to 2012. strains of influenza a/h3n2 isolated during an influenza survey from 2010 to 2012 in qinghai were enrolled by random sampling. viral rna was extracted and amplified by rt-pcr. purified pcr products were sequenced thereafter. genetic analysis of nucleic acid and the derived amino acid sequences wa ...201425118381
detection of h3n2 canine influenza virus using a quartz crystal microbalance.label-free technology-based quartz crystal microbalance (qcm) is an emerging tool in biological research. in this study, qcm was applied successfully for the rapid diagnosis of h3n2 canine influenza virus (civ) infection. prolinker™ b, a calixcrown derivative, enables antibodies to be attached to a gold-coated quartz surface and positioned in a regular pattern with the correct orientation. the prolinker-coated quartz-based assay detected h3n2 civ at lower concentrations (2(2) ha unit) than a com ...201425072379
chimeric influenza-virus-like particles containing the porcine reproductive and respiratory syndrome virus gp5 protein and the influenza virus ha and m1 proteins.both porcine reproductive and respiratory syndrome and swine influenza are acute, highly contagious swine diseases. these diseases pose severe threats for the swine industry and cause heavy economic losses worldwide. in this study, we have developed a chimeric virus-like particle (vlp) vaccine candidate for porcine reproductive and respiratory syndrome virus (prrsv) and h3n2 influenza virus and investigated its immunogenicity in mice. the ha and m1 proteins from the h3n2 influenza virus and the ...201425064513
neuraminidase gene homology contributes to the protective activity of influenza vaccines prepared from the influenza virus library.whole-virus (wv) vaccines from influenza a/duck/hokkaido/77 (h3n2), and its reassortant strains h3n4, h3n5 and h3n7, which have the same haemagglutinin (ha) gene but different neuraminidase (na) genes, were prepared from our influenza virus library. mice were intranasally immunized with equivalent doses of each vaccine (1-0.01 µg per mouse). all of the mice that received the highest dose of each vaccine (1 µg per mouse) showed equivalent high ha-inhibiting (hi) antibody titres and survived the h ...201425053564
divergent immune responses and disease outcomes in piglets immunized with inactivated and attenuated h3n2 swine influenza vaccines in the presence of maternally-derived antibodies.live-attenuated influenza virus (laiv) prime-boost vaccination previously conferred protection against heterologous h3n2 swine influenza challenge, including in piglets with maternally derived antibodies (mda). conversely, a whole-inactivated virus (wiv) vaccine was associated with enhanced disease. this study was aimed at identifying immune correlates of cross-protection. piglets with and without mda received intramuscular adjuvanted wiv or intranasal laiv, and were challenged with heterologous ...201425043588
recent h3n2 influenza virus clinical isolates rapidly acquire hemagglutinin or neuraminidase mutations when propagated for antigenic analyses.prior to serological testing, influenza viruses are typically propagated in eggs or cell culture. recent human h3n2 strains bind to cells with low avidity. here, we isolated nine primary h3n2 viral isolates from respiratory secretions of children. upon propagation in vitro, five of these isolates acquired hemagglutinin or neuraminidase mutations that increased virus binding to cell surfaces. these mutations can potentially confound serological assays commonly used to identify antigenically novel ...201424991002
performance characteristics of qualified cell lines for isolation and propagation of influenza viruses for vaccine manufacturing.cell culture is now available as a method for the production of influenza vaccines in addition to eggs. in accordance with currently accepted practice, viruses recommended as candidates for vaccine manufacture are isolated and propagated exclusively in hens' eggs prior to distribution to manufacturers. candidate vaccine viruses isolated in cell culture are not available to support vaccine manufacturing in mammalian cell bioreactors so egg-derived viruses have to be used. recently influenza a (h3 ...201424975811
influenza vaccination accelerates recovery of ferrets from lymphopenia.ferrets are a useful animal model for human influenza virus infections, since they closely mimic the pathogenesis of influenza viruses observed in humans. however, a lack of reagents, especially for flow cytometry of immune cell subsets, has limited research in this model. here we use a panel of primarily species cross-reactive antibodies to identify ferret t cells, cytotoxic t lymphocytes (ctl), b cells, and granulocytes in peripheral blood. following infection with seasonal h3n2 or h1n1pdm09 i ...201424968319
significant cross reactive antibodies to influenza virus in adults and children during a period of marked antigenic drift.little is known about the development of cross-reactive antibodies following natural exposure to pathogens. such knowledge is critical in the development of new universal influenza vaccines.201424950742
identification and genetic characterization of avian-origin h3n2 canine influenza viruses isolated from the liaoning province of china in 2012.a total of 158 serum samples and 510 nasal swab specimens were collected between september 2010 and may 2012, from dogs exhibiting respiratory symptoms, in order to investigate the epidemiology of h3n2 canine influenza viruses (civs) in the liaoning province of china. serological surveillance demonstrated that 10.8 % (17/158) of serum samples were positive for h3n2 canine influenza. two h3n2 influenza viruses, a/canine/liaoning/27/2012 and a/canine/liaoning/h6/2012, were isolated from pet dogs i ...201424928168
design, synthesis, antiviral and cytostatic activity of ω-(1h-1,2,3-triazol-1-yl)(polyhydroxy)alkylphosphonates as acyclic nucleotide analogues.the efficient synthesis of a new series of polyhydroxylated dibenzyl ω-(1h-1,2,3-triazol-1-yl)alkylphosphonates as acyclic nucleotide analogues is described starting from dibenzyl ω-azido(polyhydroxy)alkylphosphonates and selected alkynes under microwave irradiation. selected o,o-dibenzylphosphonate acyclonucleotides were transformed into the respective phosphonic acids. all compounds were evaluated in vitro for activity against a broad variety of dna and rna viruses and for cytostatic activity ...201424906510
isolation and identification of phase i metabolites of phillyrin in rats.phillyrin was one of the main chemical constituents of the fruit of forsythia suspensa (thunb.) vahl. it showed various bioactivities including antioxidant and anti-inflammatory activities. however, the metabolism of phillyrin remained unknown. this report described the isolation and identification of phase i metabolites of phillyrin in rats. nine metabolites including six new ones were isolated by various column chromatographies and high-performance liquid chromatography. their structures were ...201424879901
influenza virus pb1 and neuraminidase gene segments can cosegregate during vaccine reassortment driven by interactions in the pb1 coding region.egg-grown influenza vaccine yields are maximized by infection with a seed virus produced by "classical reassortment" of a seasonal isolate with a highly egg-adapted strain. seed viruses are selected based on a high-growth phenotype and the presence of the seasonal hemagglutinin (ha) and neuraminidase (na) surface antigens. retrospective analysis of h3n2 vaccine seed viruses indicated that, unlike other internal proteins that were predominantly derived from the high-growth parent a/puerto rico/8/ ...201424872588
influenza a virus and tlr7 activation potentiate nox2 oxidase-dependent ros production in macrophages.influenza a virus infects resident alveolar macrophages in the respiratory tract resulting in toll like receptor 7 (tlr7) activation that triggers an inflammatory response to resolve the infection. macrophages are also major sources of reactive oxygen species (ros) via the nox2-containing nadph oxidase. although ros are crucial for pathogen clearance, in response to influenza a virus, ros are touted as being culprit mediators of the lung tissue injury. the aim of the present study was to determi ...201424869957
structural basis for histone mimicry and hijacking of host proteins by influenza virus protein ns1.pathogens can interfere with vital biological processes of their host by mimicking host proteins. the ns1 protein of the influenza a h3n2 subtype possesses a histone h3k4-like sequence at its carboxyl terminus and has been reported to use this mimic to hijack host proteins. however, this mimic lacks a free n-terminus that is essential for binding to many known h3k4 readers. here we show that the double chromodomains of chd1 adopt an 'open pocket' to interact with the free n-terminal amine of h3k ...201424853335
the use of plethysmography in determining the severity of lung pathology in a mouse model of minimally lethal influenza virus infection.to characterize the impact on lung function, we assessed plethysmography parameters in a course of infection with mouse-adapted a/pennsylvania/14/2010 (h3n2) influenza virus. several parameters, represented by enhanced pause (penh) and ratio of inspiratory/expiratory time (ti/te), were observed that had early (1-7dpi) and robust changes regardless of virus challenge dose. other parameters, characterized by tidal volume (tv), breathing frequency (freq) and end inspiratory pause (eip), changed lat ...201424837607
n-glycolylneuraminic acid on human epithelial cells prevents entry of influenza a viruses that possess n-glycolylneuraminic acid binding ability.some animal influenza a viruses (iavs) bind not only to n-acetylneuraminic acid (neu5ac) but also to n-glycolylneuraminic acid (neu5gc), which has been discussed as a virus receptor. human cells cannot synthesize neu5gc due to dysfunction of the cmp-neu5ac hydroxylase (cmah) gene, which converts cmp-neu5ac to cmp-neu5gc. however, exogenous neu5gc from neu5gc-rich dietary sources is able to be metabolically incorporated into surfaces of tissue cells and may be related to enhancement of the infect ...201424829344
incidence of influenza virus infections in children in hong kong in a 3-year randomized placebo-controlled vaccine study, 2009-2012.school-aged children suffer high rates of influenza virus infections and associated illnesses each year, and are a major source of transmission in the community. however, information on the cumulative incidence of infection in specific epidemics is scarce, and there are limited studies with sufficient follow-up to identify the strength and duration of protection against reinfection.201424825868
identification of four genotypes of h3n2 swine influenza virus in pigs from southern china.in 2011, four h3n2 swine influenza viruses (sivs) were isolated from nasal swabs of four pigs (800 nasal swabs were collected from pigs showing influenza-like symptoms) in guangdong province, china. four different genotypes of h3n2 appeared among pigs in southern china, including wholly human-like h3n2 viruses, intermediate (1975) double-reassortant human h3n2 viruses (resulting from reassortment between an early human lineage and a recent human lineage), recent double-reassortant human h3n2 vir ...201424810098
novel avian influenza a (h7n9) virus induces impaired interferon responses in human dendritic cells.in march 2013 a new avian influenza a(h7n9) virus emerged in china and infected humans with a case fatality rate of over 30%. like the highly pathogenic h5n1 virus, h7n9 virus is causing severe respiratory distress syndrome in most patients. based on genetic analysis this avian influenza a virus shows to some extent adaptation to mammalian host. in the present study, we analyzed the activation of innate immune responses by this novel h7n9 influenza a virus and compared these responses to those i ...201424804732
efficient replication and strong induction of innate immune responses by h9n2 avian influenza virus in human dendritic cells.avian influenza a (h9n2) viruses have occasionally been identified in humans with upper respiratory tract infections. the novel h7n9/2013 virus identified in china shows that a low pathogenic avian influenza (lpai) virus can be highly pathogenic in humans. therefore, it is important to understand virus-host cell interactions and immune responses triggered by lpai viruses in humans. we found that lpai a/hong kong/1073/99 (h9n2) virus replicated efficiently in human dendritic cells (dcs). the h9n2 ...201425461529
continued effectiveness of laninamivir octanate hydrate for influenza treatment in japan: comparison between the 2011-2012 and 2012-2013 influenza seasons.the clinical effectiveness of laninamivir octanoate hydrate (laninamivir) was investigated in the japanese 2012-2013 influenza season for comparison with that of the japanese 2011-2012 influenza season. a total of 235 patients were enrolled, of whom 210 were evaluated for the duration of fever and other symptoms. the median durations of fever for a(h3n2) were 32.0 and 38.0 h and the median durations of symptoms for the a(h3n2) were 102.0 and 84.0 h for patients aged under 10 and 10 years or olde ...201425444677
serological survey for emerging canine h3n8 and h3n2 influenza viruses in pet and village dogs in nigeria.in nigeria, keeping of dogs as pets and guards is gaining popularity. to determine whether infection of dogs with novel canine influenza virus (civ) of equine (h3n8) and avian (h3n2) origins had occurred in nigeria, we screened pet and village dogs from lagos, ibadan, odeda and sagamu in southwestern nigeria for antibodies to civ h3n8 and h3n2.201426949788
antibody landscapes after influenza virus infection or vaccination.we introduce the antibody landscape, a method for the quantitative analysis of antibody-mediated immunity to antigenically variable pathogens, achieved by accounting for antigenic variation among pathogen strains. we generated antibody landscapes to study immune profiles covering 43 years of influenza a/h3n2 virus evolution for 69 individuals monitored for infection over 6 years and for 225 individuals pre- and postvaccination. upon infection and vaccination, titers increased broadly, including ...201425414313
the k526r substitution in viral protein pb2 enhances the effects of e627k on influenza virus replication.host-adaptive strategies, such as the e627k substitution in the pb2 protein, are critical for replication of avian influenza a viruses in mammalian hosts. here we show that mutation pb2-k526r is present in some human h7n9 influenza isolates, in nearly 80% of h5n1 human isolates from indonesia and, in conjunction with e627k, in almost all seasonal h3n2 viruses since 1970. polymerase complexes containing pb2-526r derived from h7n9, h5n1 or h3n2 viruses exhibit increased polymerase activity. pb2-52 ...201425409547
progressive antigenic drift and phylogeny of human influenza a(h3n2) virus over five consecutive seasons (2009-2013) in hangzhou, china.vaccine efficacy (ve) can be affected by progressive antigenic drift or any new reassortment of influenza viruses. to effectively track the evolution of human influenza a(h3n2) virus circulating in hangzhou, china, a total of 65 clinical specimens were selected randomly from outpatients infected by a(h3n2) viruses during the study period from november 2009 to december 2013. the results of reduced ve and antigenic drift of the correspondent epitopes (c-d-e to a-b) suggest that the current vaccine ...201425447724
substrate specificity of avian influenza h5n1 neuraminidase.to characterise neuraminidase (na) substrate specificity of avian influenza h5n1 strains from humans and birds comparing to seasonal influenza virus.201425396120
predicting evolution from the shape of genealogical trees.given a sample of genome sequences from an asexual population, can one predict its evolutionary future? here we demonstrate that the branching patterns of reconstructed genealogical trees contains information about the relative fitness of the sampled sequences and that this information can be used to predict successful strains. our approach is based on the assumption that evolution proceeds by accumulation of small effect mutations, does not require species specific input and can be applied to a ...201425385532
safety and immunogenicity of a novel cold-adapted modified-live equine influenza virus vaccine.to design and evaluate the safety and immunogenicity of a modified-live vaccine to prevent equine influenza virus (eiv) infection based on the novel reassortant cold-adapted strain a/hk/otar/6:2/2010.201425348146
epidemiological and virological characteristics of influenza viruses circulating in cambodia from 2009 to 2011.the cambodian national influenza center (nic) monitored and characterized circulating influenza strains from 2009 to 2011.201425340711
using multiple linear regression and physicochemical changes of amino acid mutations to predict antigenic variants of influenza a/h3n2 viruses.among human influenza viruses, strain a/h3n2 accounts for over a quarter of a million deaths annually. antigenic variants of these viruses often render current vaccinations ineffective and lead to repeated infections. in this study, a computational model was developed to predict antigenic variants of the a/h3n2 strain. first, 18 critical antigenic amino acids in the hemagglutinin (ha) protein were recognized using a scoring method combining phi (ϕ) coefficient and information entropy. next, a pr ...201425227088
enhanced natural killer-cell and t-cell responses to influenza a virus during pregnancy.pregnant women experience increased morbidity and mortality after influenza infection, for reasons that are not understood. although some data suggest that natural killer (nk)- and t-cell responses are suppressed during pregnancy, influenza-specific responses have not been previously evaluated. thus, we analyzed the responses of women that were pregnant (n = 21) versus those that were not (n = 29) immediately before inactivated influenza vaccination (iiv), 7 d after vaccination, and 6 wk postpar ...201425246558
emergence of truncated pb1-f2 protein of h3n2 influenza virus during its epidemic period in jiangsu province, china.pb1-f2 protein has been proven to increase the pathogenicity of influenza a virus (iav) strains in primary infection and in secondary bacterial infection. it can also regulate the activity of viral polymerase. however, it was shown in another retrospective study that a portion of iavs do not express full-length pb1-f2 protein during virus development; different kinds of stop codons cause exits in the open reading frames and form pb1-f2 gene products with the corresponding genotypes. truncated pb ...201424762594
akt inhibitor mk2206 prevents influenza ph1n1 virus infection in vitro.the influenza ph1n1 virus caused a global flu pandemic in 2009 and continues manifestation as a seasonal virus. better understanding of the virus-host cell interaction could result in development of better prevention and treatment options. here we show that the akt inhibitor mk2206 blocks influenza ph1n1 virus infection in vitro. in particular, at noncytotoxic concentrations, mk2206 alters akt signaling and inhibits endocytic uptake of the virus. interestingly, mk2206 is unable to inhibit h3n2, ...201424752266
influenza a subtype h3 viruses in feral swine, united states, 2011-2012.to determine whether, and to what extent, influenza a subtype h3 viruses were present in feral swine in the united states, we conducted serologic and virologic surveillance during october 2011-september 2012. these animals were periodically exposed to and infected with a(h3n2) viruses, suggesting they may threaten human and animal health.201424751326
development of a reverse transcription loop-mediated isothermal amplification assay for the rapid diagnosis of avian influenza a (h7n9) virus infection.a genetic diagnosis system for detecting avian influenza a (h7n9) virus infection using reverse transcription-loop-mediated isothermal amplification (rt-lamp) technology was developed. the rt-lamp assay showed no cross-reactivity with seasonal influenza a (h3n2 and h1n1pdm09) or influenza b viruses circulating in humans or with avian influenza a (h5n1) viruses. the sensitivity of the rt-lamp assay was 42.47 copies/reaction. considering the high specificity and sensitivity of the assay for detect ...201424747008
intrahost dynamics of influenza virus reassortment.the segmented nature of the influenza virus genome allows reassortment between coinfecting viruses. this process of genetic exchange vastly increases the diversity of circulating influenza viruses. the importance of reassortment to public health is clear from its role in the emergence of a number of epidemiologically important viruses, including novel pandemic and epidemic strains. to gauge its impact on within-host genomic variation, we tracked reassortment in coinfected guinea pigs over time a ...201424741099
analysis of the acute-phase protein response in pigs to clinical and subclinical infection with h3n2 swine influenza virus.swine influenza (si) is a contagious, important respiratory disease. diagnosis of si is based on the clinical signs, confirmed by the detection of viral rna or specific antibodies. however, the infection is much more frequent than the disease.201424734294
duration of the protective immune response after prime and booster vaccination of yearlings with a live modified cold-adapted viral vaccine against equine influenza.we previously created a live vaccine against equine influenza based the new reassortant cold-adapted (ca) strain a/hk/otar/6:2/2010. the live vaccine contains surface proteins (ha, na) from the wild-type virus a/equine/otar/764/2007 (н3n8; american lineage florida clade 2), and internal proteins (pb2, pb1, pa, np, m, ns) from the attenuated ca donor virus a/hong kong/1/68/162/35ca (h3n2). to determine the safety and duration of the protective immune responses, 90 yearlings were intranasally vacc ...201424726250
conserved neutralizing epitope at globular head of hemagglutinin in h3n2 influenza viruses.neutralizing antibodies that target the hemagglutinin of influenza virus either inhibit binding of hemagglutinin to cellular receptors or prevent the low-ph-induced conformational change in hemagglutinin required for membrane fusion. in general, the former type of antibody binds to the globular head formed by ha1 and has narrow strain specificity, while the latter type binds to the stem mainly formed by ha2 and has broad strain specificity. in the present study, we analyzed the epitope and funct ...201424719430
receptor mimicry by antibody f045-092 facilitates universal binding to the h3 subtype of influenza virus.influenza viruses present a significant health challenge each year, as in the h3n2 epidemic of 2012-2013. here we describe an antibody, f045-092, that possesses broadly neutralizing activity against the entire h3 subtype and accommodates the natural variation and additional glycosylation in all strains tested from 1963 to 2011. crystal structures of f045-092 in complex with has from 1975 and 2011 h3n2 viruses reveal the structural basis for its neutralization breadth through insertion of its 23- ...201424717798
a broadly neutralizing human monoclonal antibody directed against a novel conserved epitope on the influenza virus h3 hemagglutinin globular head.most neutralizing antibodies elicited during influenza virus infection or vaccination target immunodominant, variable epitopes on the globular head region of hemagglutinin (ha), which leads to narrow strain protection. in this report, we describe the properties of a unique anti-ha monoclonal antibody (mab), d1-8, that was derived from human b cells and exhibits potent, broad neutralizing activity across antigenically diverse influenza h3 subtype viruses. based on selection of escape variants, we ...201424696468
full-genome sequences of influenza h3n2 virus strains isolated from finnish patients during the 2012-2013 epidemic season.here, we sequenced 10 influenza a(h3n2) virus genomes isolated from finnish patients diagnosed with flu-like illness during the 2012-2013 influenza season. the alignment showed a high number of amino acid substitutions (238 in total) in only 10 samples, proving that a high mutation rate exists in seasonal influenza a viruses.201424675845
a zanamivir dimer with prophylactic and enhanced therapeutic activity against influenza viruses.emerging drug resistance to antiviral therapies is an increasing challenge for the treatment of influenza virus infections. one new antiviral compound, bta938, a dimeric derivative of the viral neuraminidase inhibitor zanamivir, contains a 14-carbon linker bridging two zanamivir moieties. in these studies, we evaluated antiviral efficacy in cell cultures infected with influenza virus and in mouse models of lethal influenza using h1n1pdm09, h3n2 and oseltamivir-resistant (h275y) viruses.201424777908
recombinant porcine surfactant protein d inhibits influenza a virus replication ex vivo.influenza is a major burden to public health. due to high mutation rates and selection pressure, mutant viruses emerge which are resistant to currently used antiviral drugs. therefore, there is a need for the development of novel classes of antiviral drugs that suffer less from the emergence of resistant viruses. antiviral drugs based on collectin-like surfactant protein d (sp-d) may fulfil these requirements. especially porcine sp-d displays strong antiviral activity to influenza a viruses. in ...201424389095
structural insights into the membrane fusion mechanism mediated by influenza virus hemagglutinin.membrane fusion is involved in many fundamental cellular processes and entry of enveloped viruses into host cells. influenza type a virus ha has long served as a paradigm for mechanistic studies of protein-mediated membrane fusion via large-scale structural rearrangements induced by acidic ph. here we report the newly determined crystal structure of influenza b virus ha2 in the postfusion state. together with a large number of previously determined prefusion structures of influenza a and b virus ...201424433110
repetitive fulminant influenza myocarditis requiring the use of circulatory assist devices.a 52-year-old man was admitted to our hospital due to shortness of breath that developed one week after the diagnosis of influenza infection. he had a past history of myocarditis associated with influenza b infection 16 years before the current admission. the patient's left ventricular function showed diffuse hypokinesis with a left ventricular ejection fraction of 28%. due to the progression of heart failure, the infusion of catecholamines and insertion of an intra-aortic balloon pump were requ ...201424429449
novel oral anti-influenza prodrug candidate av5075s.development of a novel drug candidate with improved activity against influenza virus neuraminidase (na) compared with currently available therapeutics.201424428978
mechanism of a decrease in potency for the recombinant influenza a virus hemagglutinin h3 antigen during storage.the recombinant hemagglutinin (rha)-based influenza vaccine flublok® has recently been approved in the united states as an alternative to the traditional egg-derived flu vaccines. flublok is a purified vaccine with a hemagglutinin content that is threefold higher than standard inactivated influenza vaccines. when rha derived from an h3n2 influenza virus was expressed, purified, and stored for 1 month, a rapid loss of in vitro potency (∼50%) was observed as measured by the single radial immunodif ...201424425059
intrasubtype reassortments cause adaptive amino acid replacements in h3n2 influenza genes.reassortments and point mutations are two major contributors to diversity of influenza a virus; however, the link between these two processes is unclear. it has been suggested that reassortments provoke a temporary increase in the rate of amino acid changes as the viral proteins adapt to new genetic environment, but this phenomenon has not been studied systematically. here, we use a phylogenetic approach to infer the reassortment events between the 8 segments of influenza a h3n2 virus since its ...201424415946
reserve autophagic capacity in alveolar epithelia provides a replicative niche for influenza a virus.autophagy contributes to cellular homeostasis through metabolite recycling and degradation of cytotoxic protein aggregates and damaged organelles. although recent studies have established that the requirement for basal autophagy is largely tissue specific, the importance of autophagy for alveolar epithelial cell homeostasis remains an important knowledge gap. in the present study we generated two mouse models, with > 90% or > 50% recombination at the atg5 locus in the distal respiratory epitheli ...201424661119
cluster of influenza a cases in vaccinated population of adults in virology laboratory in glasgow in december 2012.the majority of influenza infections during the 2012/2013 influenza season in scotland have been due to influenza a h3n2. we report an outbreak of influenza a h3n2 in a vaccinated population of adults in the regional virology laboratory in glasgow. this investigation was carried out to confirm the epidemiological link between cases.201424659378
emergence of g186d mutation in the presence of r292k mutation in an immunocompromised child infected with influenza a/h3n2 virus, treated with oseltamivir.an immunocompromised child with influenza a/h3n2 virus infection, treated with oseltamivir from day 1, had nasal swabs taken on days 1, 4, 7, and 10 of the illness. pyrosequencing showed increasing proportions of viruses with r292k (neuraminidase gene) and g186d (hemagglutinin gene) mutations, resulting in a viral load rebound by day 10.201424599981
a phospha-oseltamivir-biotin conjugate as a strong and selective adhesive for the influenza virus.we present the synthesis and application of a molecule containing both the powerful influenza neuraminidase (na) inhibitor phospha-oseltamivir and d-biotin, connected via an undecaethylene glycol spacer. it inhibits influenza virus neuraminidase (from the h3n2 x31 virus) in the same range as oseltamivir, with a slow off-rate, and produces a stable na-coated surface when loaded onto streptavidin-coated biosensors. purified x31 virus binds to these loaded biosensors with an apparent dissociation c ...201424594352
(19)f nmr reveals multiple conformations at the dimer interface of the nonstructural protein 1 effector domain from influenza a virus.nonstructural protein 1 of influenza a virus (ns1a) is a conserved virulence factor comprised of an n-terminal double-stranded rna (dsrna)-binding domain and a multifunctional c-terminal effector domain (ed), each of which can independently form symmetric homodimers. here we apply (19)f nmr to ns1a from influenza a/udorn/307/1972 virus (h3n2) labeled with 5-fluorotryptophan, and we demonstrate that the (19)f signal of trp187 is a sensitive, direct monitor of the ed helix:helix dimer interface. ( ...201424582435
a predictive fitness model for influenza.the seasonal human influenza a/h3n2 virus undergoes rapid evolution, which produces significant year-to-year sequence turnover in the population of circulating strains. adaptive mutations respond to human immune challenge and occur primarily in antigenic epitopes, the antibody-binding domains of the viral surface protein haemagglutinin. here we develop a fitness model for haemagglutinin that predicts the evolution of the viral population from one year to the next. two factors are shown to determ ...201424572367
live attenuated influenza vaccine enhances colonization of streptococcus pneumoniae and staphylococcus aureus in mice.community interactions at mucosal surfaces between viruses, like influenza virus, and respiratory bacterial pathogens are important contributors toward pathogenesis of bacterial disease. what has not been considered is the natural extension of these interactions to live attenuated immunizations, and in particular, live attenuated influenza vaccines (laivs). using a mouse-adapted laiv against influenza a (h3n2) virus carrying the same mutations as the human flumist vaccine, we find that laiv vacc ...201424549845
substitutions near the hemagglutinin receptor-binding site determine the antigenic evolution of influenza a h3n2 viruses in u.s. swine.swine influenza a virus is an endemic and economically important pathogen in pigs, with the potential to infect other host species. the hemagglutinin (ha) protein is the primary target of protective immune responses and the major component in swine influenza a vaccines. however, as a result of antigenic drift, vaccine strains must be regularly updated to reflect currently circulating strains. characterizing the cross-reactivity between strains in pigs and seasonal influenza virus strains in huma ...201424522915
the duration of fever and other symptoms after the initiation of laninamivir octanoate hydrate in the japanese 2011-2012 influenza season.laninamivir octanoate hydrate (laninamivir) is a long-acting, single inhalation neuraminidase inhibitor that was approved in japan in 2010 for the treatment of influenza a and b virus infection. we investigated the duration of fever and other symptoms after the initiation of laninamivir in the japanese 2011-2012 influenza season. virus isolation was done and the 50% inhibitory concentration (ic₅₀) was measured for the virus isolates at days 1 and 5. for 211 patients (a(h3n2): 190, b: 21), the me ...201424486167
intranasal vaccination promotes detrimental th17-mediated immunity against influenza infection.influenza disease is a global health issue that causes significant morbidity and mortality through seasonal epidemics. currently, inactivated influenza virus vaccines given intramuscularly or live attenuated influenza virus vaccines administered intranasally are the only approved options for vaccination against influenza virus in humans. we evaluated the efficacy of a synthetic toll-like receptor 4 agonist crx-601 as an adjuvant for enhancing vaccine-induced protection against influenza infectio ...201424465206
inferring the antigenic epitopes for highly pathogenic avian influenza h5n1 viruses.the evasion of influenza virus from host immune surveillance is mainly mediated through its surface protein hemagglutinin (ha), the main component of influenza vaccine. thus, identification of influenza virus antigenic epitopes on ha can not only help us understand the molecular mechanisms of viral immune escape but also facilitate vaccine strain selection. despite previous efforts, there is a lack of systematic definition of the antigenic epitopes for the highly pathogenic avian influenza (hpai ...201424380681
exploration of risk factors contributing to the presence of influenza a virus in swine at agricultural fairs.influenza a virus infections occurring in exhibition swine populations at agricultural fairs during 2012 served as a source of h3n2 variant influenza a viruses transmitted to humans resulting in more than 300 documented cases. prior to the outbreak, this investigation was initiated to identify fair-level risk factors contributing to influenza a virus infections in pigs at agricultural fairs. as part of an ongoing active surveillance program, nasal swabs and associated fair-level metadata were co ...201426038494
genomewide analysis of reassortment and evolution of human influenza a(h3n2) viruses circulating between 1968 and 2011.influenza a(h3n2) viruses became widespread in humans during the 1968 h3n2 virus pandemic and have been a major cause of influenza epidemics ever since. these viruses evolve continuously by reassortment and genomic evolution. antigenic drift is the cause for the need to update influenza vaccines frequently. using two data sets that span the entire period of circulation of human influenza a(h3n2) viruses, it was shown that influenza a(h3n2) virus evolution can be mapped to 13 antigenic clusters. ...201424371052
predicting the emergence of h3n2 influenza viruses reveals contrasted modes of evolution of ha and na antigens.vaccine design for rapidly changing viruses is based on empirical surveillance of strains circulating in a given season to assess those that will most likely spread during the next season. the choice of which strains to include in the vaccine is critical, as an erroneous decision can lead to a nonimmunized human population that will then be at risk in the face of an epidemic or, worse, a pandemic. here, we present the first steps toward a very general phylogenetic approach to predict the emergen ...201424343641
a common solution to group 2 influenza virus neutralization.the discovery and characterization of broadly neutralizing antibodies (bnabs) against influenza viruses have raised hopes for the development of monoclonal antibody (mab)-based immunotherapy and the design of universal influenza vaccines. only one human bnab (cr8020) specifically recognizing group 2 influenza a viruses has been previously characterized that binds to a highly conserved epitope at the base of the hemagglutinin (ha) stem and has neutralizing activity against h3, h7, and h10 viruses ...201424335589
comparative analysis of micrornas from the lungs and trachea of dogs (canis familiaris) infected with canine influenza virus.micrornas (mirnas) are a class of endogenous non-coding small rnas of 18-22-nucleotides in length that regulate gene expression at the post-transcriptional level. the objective of this study was to examine the differences in the mirna expression profiles of the lungs and trachea of beagle dogs infected with canine influenza virus (civ). total rna was isolated from lung and trachea tissues of beagle dogs infected and non-infected with h3n2 civ at 4 dpi. a total of 41,512,315 and 39,107,475 reads ...201424333372
genetic characterization of seasonal influenza a (h3n2) viruses in ontario during 2010-2011 influenza season: high prevalence of mutations at antigenic sites.the direct effect of antigenic site mutations in influenza viruses on antigenic drift and vaccine effectiveness is poorly understood.201424313991
comparative analysis of receptor-binding specificity and pathogenicity in natural reassortant and non-reassortant h3n2 swine influenza virus.genetic reassortment between human and avian influenza viruses can create pandemic viruses. influenza surveillance of pigs in jilin province, in china during 2007-2008 revealed that there were two distinguishable genotypes: a human-like h3n2 genotype and a double-reassortant genotype derived from the human h3n2 and avian h5 viruses. in this study, viral infection potential, replication kinetics, and pathogenicity were compared. the solid-phase binding assay demonstrated that both viruses promine ...201424305615
influenza vaccine response in adults exposed to perfluorooctanoate and perfluorooctanesulfonate.supported by several epidemiological studies and a large number of animal studies, certain polyfluorinated alkyl acids are believed to be immunotoxic, affecting particularly humoral immunity. our aim was to investigate the relationship between the antibody response following vaccination with an inactivated trivalent influenza vaccine and circulating levels of perfluorooctanoate (pfoa) and perfluorooctanesulfonate (pfos). the study population consisted of 411 adults living in the mid-ohio region ...201424284791
synthesis of 1,2,3-triazolyl nucleoside analogs as potential anti-influenza a (h3n2 subtype) virus agents.montmorillonite k10 impregnated with copper dichloride and potassium iodide (cucl2 /ki/k10) was used as catalyst in the cycloaddition of azides and propargylnucleobases, to provide the corresponding 1,4-disubstituted 1,2,3-triazoles in good yield. all compounds 16-23 were evaluated for their antiviral activity in vitro. compound 18 showed moderate inhibition against influenza virus a (h3n2).201424272912
recombinant influenza a h3n2 viruses with mutations of ha transmembrane cysteines exhibited altered virological characteristics.influenza a h3n2 virus as the cause of 1968 pandemic has since been circulating in human and swine. our earlier study has shown that mutations of one or two cysteines in the transmembrane domain of h3 hemagglutinin (ha) affected the thermal stability and fusion activity of recombinant ha proteins. here, we report the successful generation of three recombinant h3n2 mutant viruses (c540s, c544l, and 2c/sl) with mutations of one or two transmembrane cysteines of ha in the background of a/swine/guan ...201424272698
novel h7n9 influenza virus shows low infectious dose, high growth rate, and efficient contact transmission in the guinea pig model.the zoonotic outbreak of h7n9 subtype avian influenza virus that occurred in eastern china in the spring of 2013 resulted in 135 confirmed human cases, 44 of which were lethal. sequencing of the viral genome revealed a number of molecular signatures associated with virulence or transmission in mammals. we report here that, in the guinea pig model, a human isolate of novel h7n9 influenza virus, a/anhui/1/2013 (an/13), is highly dissimilar to an h7n1 avian isolate and instead behaves similarly to ...201424227867
the effect of the mdck cell selected neuraminidase d151g mutation on the drug susceptibility assessment of influenza a(h3n2) viruses.propagation of influenza a(h3n2) viruses in mdck cells has been associated with the emergence of neuraminidase (na) variants carrying a change at residue 151. in this study, the pyrosequencing assay revealed that ∼90% of a(h3n2) virus isolates analyzed (n=150) contained more than one amino acid variant (d/g/n) at position 151. susceptibilities of the virus isolates to zanamivir and oseltamivir were assessed using the chemiluminescent and fluorescent na inhibition (ni) assays. in the chemilumines ...201424239666
loss and gain of n-linked glycosylation sites in globular head and stem of ha found in a/h3n2 flu fatal and severe cases during 2013 tunisia flu seasonal survey.glycosylation on the globular head of the hemagglutinin (ha) protein of influenza virus acts as an important target for recognition and destruction of virus by innate immune proteins of the collectin family. in the current study, we have characterized the dynamic amino acid changes at n-linked glycosylation sites of full length sequences of ha genes of 5 a/h3n2 tunisian strains isolates from mild, severe, and fatal cases. compared to the reference strain, a/perth/16/2009 substitutions in potenti ...201424174280
genetic characterization of canine influenza a virus (h3n2) in thailand.in january 2012, several clinical cases of dogs with flu-like symptoms, including coughing, sneezing, nasal discharge, and fever, were reported in a small-animal hospital located in bangkok, thailand. one influenza a virus was identified and characterized as an avian-like influenza virus h3n2. the virus was named a/canine/thailand/cu-dc5299/12. a phylogenetic analysis indicated that the canine virus belonged to an avian eurasian lineage and was genetically related to the canine influenza viruses ...201424101341
rapid strategy for screening by pyrosequencing of influenza virus reassortants--candidates for live attenuated vaccines.live attenuated influenza vaccine viruses (laivs) can be generated by classical reassortment of gene segments between a cold adapted, temperature sensitive and attenuated master donor virus (mdv) and a seasonal wild-type (wt) virus. the vaccine candidates contain hemagglutinin (ha) and neuraminidase (na) genes derived from the circulating wt viruses and the remaining six genes derived from the mdv strains. rapid, efficient selection of the viruses with 6∶2 genome compositions from the large numb ...201424647786
detection of influenza a virus nucleoprotein antibodies in oral fluid specimens from pigs infected under experimental conditions using a blocking elisa.in commercial swine populations, influenza is an important component of the porcine respiratory disease complex (prdc) and a pathogen with major economic impact. previously, a commercial blocking elisa (flockchek(™) avian influenza virus multis-screen(®) antibody test kit, idexx laboratories, inc., westbrook, me, usa) designed to detect influenza a nucleoprotein (np) antibodies in avian serum was shown to accurately detect np antibodies in swine serum. the purpose of this study was to determine ...201423046061
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